Ferns display an elevated degree of phenotypic plasticity to changes in irradiance levels; however, only a few reports deal with their response to different light conditions. To get an insight into the extent of phenotypic plasticity of the fern Phyllitis scolopendrium, thriving in a forested area along a radiation gradient at the entrance of a cave, we examined selected biochemical, morphological, and physiological frond traits of the ferns from three different habitats. Sampling was performed two times during the vegetation season, in April and June. We also measured frond optical properties to point out the differences in leaf/light interactions between different plant samples. According to frond size, the middle habitat, receiving 125 µmol m−2s−1 of photosynthetically active radiation at both sampling times, appeared to be the most favourable. The production of UV-absorbing substances was highest in the habitat with the lowest radiation level. At the beginning of the season, the level of photosynthetic pigments in this habitat was the same as in the other habitats, while it was significantly lower in June when the tree canopy was closed. Frond reflectance was similar when comparing habitats and different sampling times. The most significant differences were obtained in the UV-A and near-infrared regions. The reflectance spectra depended mainly on frond biochemical properties, which altogether explained 54% (p ≤ 0.05) of the spectra variability. Frond transmittance depended on both, morphological parameters, explaining 51% (p ≤ 0.05), and frond biochemistry, explaining 73% (p ≤ 0.05) of the spectra variability. P. scolopendrium was revealed to be highly plastic regarding light conditions. The shapes of the frond reflectance and transmittance optical curves were similar to those typical of leaves of seed plants. The fronds exhibited high morphological plasticity when comparing different habitats. However, their biochemical and optical traits differed more between the two sampling times than between the habitats.